ES2440390A1 - Compositions comprising hyaluronic acid and dermatan sulphate for the treatment of being overweight and obesity - Google Patents

Abstract

The present invention relates to a composition comprising hyaluronic acid and dermatan sulphate for use in the treatment or prevention of overweight, obesity, insulin resistance, type 2 diabetes, fatty liver or dyslipidemia or for use in reducing food intake, inducing the feeling of satiety, reducing appetite, reducing body weight, preventing weight gain, reducing body fat, reducing fat formation, reducing blood cholesterol, maintaining normal blood cholesterol levels, reducing blood triglycerides, maintaining normal blood triglyceride levels or increasing insulin sensitivity. The composition can further contain collagen hydrolysate. The composition can be in the form of a pharmaceutical composition, a food, a functional food, a medical food or a food supplement.

Description

Compositions for the treatment of overweight and obesity

Technical sector of the invention

The present invention relates to compositions for use in the treatment, prevention or reduction of overweight or obesity, as well as of diseases, conditions or metabolic disorders related to overweight or obesity and together with which they make up the metabolic syndrome, such as insulin resistance, type 2 diabetes, fatty liver or dyslipidemia, the compositions being in the form of pharmaceutical compositions.

Prior art

Obesity, defined as excess body weight in the form of fat, is an important risk factor for serious clinical disorders, including type 2 diabetes and cardiovascular disease, and is a problem of increasing incidence and difficult treatment that entails a significant socio-health expenditure (see http://www.iaso.org/iotf/obesity/). It is known that many people who are overweight or obese develop insulin resistance and hyperinsulinemia, glucose intolerance, dyslipidemia, and hypertension, characteristics that together make up the so-called metabolic syndrome.

One of the most frequent metabolic complications in obesity is insulin resistance (A. Astrup &

N. Finer, Obes. Rev. 1, 57-59 (2000)). The connection between both conditions has been extensively studied and seems to be due to the fact that, in obese individuals, adipose tissue releases increased or abnormal amounts of non-esterified fatty acids, hormones (adipokines) and pro-inflammatory cytokines, among other factors, which favor Loss of insulin sensitivity in different tissues (SE Kahn et al., Nature 444, 840-846 (2006)). Resistin, in particular, is a protein secreted by adipocytes and / or macrophages infiltrated in obese adipose tissue that acts as a pro-inflammatory factor and insulin resistance, and whose circulating levels are elevated in obese individuals (DR Schwartz & MA Lazar, Trends Endocrinol, Metab. 22, 259-265 (2011)). Resistin reduction correlates with increases in insulin sensitivity

(C.M. Steppan et al., Nature 409, 307-312 (2001); F. Felipe et al., Diabetes 53, 882-889 (2004)).

Insulin resistance is clinically relevant because it is closely associated with type 2 diabetes and other problems such as hypertension, dyslipidemia, and blood clotting defects and fibrinolysis, all of which are in turn related to cardiovascular disease (G Boden, Curr. Opin. Endocrinol. Diabetes Obes. 18, 139-143 (2011)). In general, it is accepted that the primary defect in the development of type 2 diabetes is the deterioration of insulin sensitivity, which is followed by compensatory hyperinsulinemia, and hyperglycemia when compensation mechanisms fail. Therefore, the improvement of glucose-insulin homeostasis is a health objective of specific interest in itself.

Strategies for weight reduction based solely on caloric restriction and increased physical activity are difficult to follow, and have been ineffective against the current obesity pandemic. At the same time, there is evidence that specific nutrients and other food ingredients influence biochemical processes that impact on energy balance and fat accumulation so that they can favor a leaner phenotype (KH Kim & Y. Park, Annu. Rev. Food Sci Technol. 2, 237-257 (2011);

C. Pico et al., Rev. Esp. Obes. 4, 156-174 (2006); E. M. Kovacs & D. J. Mela, Obes. Rev. 7, 59-78 (2006)). This evidence is the basis for the development of food supplements or functional foods for the control of obesity.

The following main strategies in functional feeding for the control of obesity have been defined: (i) reduction of intake, via modulation of feelings of hunger and / or satiety, or limiting the bioavailability of nutrients; (ii) reduction of caloric density in light foods; (iii) stimulation of energy expenditure through activation of adaptive thermogenesis; and (iv) redirection of nutrients to tissues and metabolic pathways that consume them, thereby favoring fat deposition (C. Pico et al., Rev. Esp. Obes. 4, 156-174 (2006)). The use of nutrients or combinations of nutrients capable of simultaneously affecting several of these processes is considered of special interest (C Pico et al., Rev. Esp. Obes. 4, 156-174 (2006)).

Inhibition of adipogenesis is another possible anti-obesity strategy (K. H. Kim & Y. Park, Annu. Rev. Food Sci. Technol. 2, 237-257 (2011)). Adipogenesis is the process of differentiating adipocytes from precursor cells, and has been widely studied mainly in cellular models (B. Feve, Best Pract. Res. Clin. Endocrinol. Metab. 19, 483-499B (2005)) . In humans, a considerable percentage of adipocytes, around 10%, are renewed annually throughout adulthood by coordinating de novo adipogenesis and death of pre-existing adipocytes (KL Spalding et al., Nature 453, 783-787 ( 2008)). Many obese individuals have an excessive number of adipocytes (hyperplasia) in their adipose deposits, and these individuals are especially refractory to long-term weight loss and prone to the known yo-yo effect. The pharmacological or nutritional control of adipogenesis emerges in this context as a new therapeutic target in the control of obesity, as an adjunct in conventional strategies of control of energy balance: inhibition of adipogenesis can help to normalize (reduce) the number of adipocytes, and with it the maintenance of weight after weight loss (P. Arner & KL Spalding, Biochem. Biophys. Res. Commun. 396, 101-104 (2010)).

Glycosaminoglycans (GAGs) are polymeric biomolecules of high molecular weight formed by repeats of a disaccharide unit. They are found primarily in living organisms, where they develop different physiological functions.

Hyaluronic acid is a non-sulfated glycosaminoglycan, with a polymeric structure characterized by a repeating disaccharide, consisting of N-acetyl-D-glucosamine monosaccharides and D-glucuronic acid. It is one of the main components of cartilage, synovial membrane and synovial fluid. In particular, its use is important in the treatment of joint dysfunctions such as osteoarthritis, usually intra-articularly. Its use has also been described in ophthalmology, wound healing, as well as cosmetics.

Dermatan sulfate is a sulfated glycosaminoglycan with a polymeric structure consisting mostly of disaccharides of N-acetyl-D-galactosamine sulfated in position 4 and L-iduronic acid, often sulfated in position 2. It has been described that it participates in the repair of wounds and in the regulation of blood coagulation. This substance is used largely in cosmetics.

The collagen hydrolyzate consists of a mixture of amino acids and low molecular weight peptides. It is obtained by controlled enzymatic hydrolysis of the collagenous protein contained in the skin and other connective tissues. It is mostly used in cosmetics.

US patent application 2005084518 describes a health food containing hyaluronic acid and dermatan sulfate. Said food is useful for beautifying the skin.

US 7,763,594 describes the use of a hyaluronic acid and dermatan sulfate composition to treat osteoarthritis.

In view of the above, providing compositions for the treatment, prevention or control of overweight and / or obesity, as well as some of its metabolic complications, is a subject of great interest in the therapeutic field.

Explanation of the invention.

The present inventors have found that the compositions of the present invention, defined below, exert a synergistic anti-adipogenic effect, even in the presence of a pro-adipogenic hormonal cocktail; inhibit the expression of resistin (adipokine related to insulin resistance); reduce weight gain and accumulated energy intake in mice fed ad libitum; reduce insulinemia and triglyceridemia in mice; and cause weight and fat loss, as well as fat mass gain, in obese mice. Therefore, said compositions can be used in the treatment or prevention of overweight, obesity, insulin resistance, type 2 diabetes, fatty liver or dyslipidemia.

The present invention relates to the use of a composition comprising hyaluronic acid and dermatan sulfate for the preparation of a medicament for the treatment or prevention of at least one of the diseases that make up the metabolic syndrome in a mammal, selected from the group consisting of overweight, obesity, insulin resistance, type 2 diabetes, fatty liver and dyslipidemia.

Preferably, the use of said composition is for the preparation of a medicament for the treatment

or prevention of overweight associated with metabolic syndrome in a mammal. Equally preferred is the use of said composition for the preparation of a medicament for the treatment or prevention of obesity associated with metabolic syndrome in a mammal. Equally preferred is the use of said composition for the preparation of a medicament for the treatment or prevention of insulin resistance associated with metabolic syndrome in a mammal. Equally preferred is the use of said composition for the preparation of a medicament for the treatment or prevention of type 2 diabetes associated with metabolic syndrome in a mammal. Equally preferred is the use of said composition for the preparation of a medicament for the treatment or prevention of fatty liver associated with metabolic syndrome in a mammal. Equally preferred is the use of said composition for the preparation of a medicament for the treatment or prevention of dyslipidemia associated with the metabolic syndrome in a mammal, and particularly the case in which the dyslipidemia is hypercholesterolemia, hypertriglyceridemia or both at the same time. , that is, hypercholesterolemia and hypertriglyceridemia.

In a preferred embodiment, insulin resistance is associated with overweight or obesity, type 2 diabetes is associated with overweight or obesity, fatty liver is associated with overweight or obesity or dyslipidemia is associated with overweight or obesity , said overweight or obesity associated with the metabolic syndrome in a mammal.

In another preferred embodiment, hyaluronic acid and dermatan sulfate are present in a weight ratio of hyaluronic acid to dermatan sulfate between 1: 0.05 and 1: 0.75, preferably between 1: 0.10 and 1: 0.50. The weight ratio especially hyaluronic acid to dermatan sulfate is 1: 0.25.

In another equally preferred embodiment, the composition further comprises collagen hydrolyzate. Preferably, hyaluronic acid, dermatan sulfate and collagen hydrolyzate are present in a weight ratio of hyaluronic acid to dermatan sulfate to collagen hydrolyzate between 1: 0.06: 0.03 and 1: 0.80: 0.40, preferably between 1: 0.10: 0.05 and 1: 0.40: 0.20. The weight ratio of hyaluronic acid to dermatan sulfate to especially preferred collagen hydrolyzate is 1: 0.20: 0.10.

In another equally preferred embodiment the mammal is a human.

Preferably, the composition or medicament is adapted for oral or parenteral administration. The most preferred is oral administration.

In the present invention, the term "comprises" should be construed so that it also includes the case of "consists solely of" and that of "consists essentially of".

In the present invention, when talking about hyaluronic acid and dermatan sulfate, any salt thereof is included, for example sodium, potassium or calcium salt.

Preferably, the hyaluronic acid and dermatan sulfate of the compositions of the present invention are in the form of sodium salt.

In the present invention, the average molecular weights of hyaluronic acid, dermatan sulfate and chondroitin sulfate have been determined by gel permeation chromatography (GPC).

In the present invention, the abbreviation HA means hyaluronic acid (Hyaluronic Acid), the abbreviation DS means dermatan sulfate (Dermatan Sulphate), the abbreviation CH stands for collagen hydrolyzate (Collagen Hydrolysate) and the abbreviation CS means chondroitin sulfate (Chondroitin Sulphate ).

In the present invention the following terms have the indicated meaning:

"Metabolic syndrome" refers to when several of a number of different risk factors, such as insulin resistance and hyperinsulinemia, glucose intolerance, dyslipidemia, and hypertension are associated in the same person with overweight or obesity.

"Inducing the feeling of satiety" refers to inducing the body (the individual) to feel satisfied and not hungry, understood as the need for immediate food intake.

"Maintain normal blood cholesterol levels" refers to keeping total cholesterol levels below 200 mg / dL, keeping LDL cholesterol levels below 100 mg / dL and keeping HDL cholesterol levels above 40 mg / dL

"LDL cholesterol" refers to cholesterol linked to low density lipoproteins (Low Density Lipoproteins), also known as "bad cholesterol."

"HDL cholesterol" refers to cholesterol linked to high density lipoproteins (High Density Lipoproteins), also known as "good cholesterol."

"Hypercholesterolemia" refers to when the levels of total cholesterol and / or LDL cholesterol in the blood are elevated.

"Maintaining normal blood triglyceride levels" refers to maintaining triglyceride levels below 150 mg / dL.

"Parenteral route" refers to when medications or nutrients are introduced into the body without being absorbed by the gastro-intestinal tract. The most frequent parentelar pathways are intravenous, subcutaneous and intramuscular.

The hyaluronic acid and dermatan sulfate of the compositions of the present invention can be obtained by extractive procedures from bird or mammalian tissues or by biotechnology.

The average molecular weights of hyaluronic acid and dermatan sulfate of the compositions of the present invention may vary depending on the method of production. Preferably, the average molecular weight of hyaluronic acid is between 300,000 and 2,000,000 daltons, more preferably between 800,000 and 1,000,000 daltons and that of dermatan sulfate is between 10,000 and 50,000 daltons.

The hyaluronic acid of the compositions of the present invention can be obtained, by extraction, from tissues of birds or mammals, for example from vitreous humor, mammalian skin, umbilical cord

or bird crests or by fermentation of microorganisms (for example Streptococcus), following procedures described in the literature (D.A. Swann, Biochim. Biophys. Acta 156, 17-30 (1968); US Patent 4,780,414).

Preferably, the hyaluronic acid of the compositions of the present invention is a commercial product available at www.bioiberica.com. It is obtained from rooster crests, which once chopped are digested with a proteolytic enzyme. The enzyme is subsequently deactivated by heating, filtered, the dermatan sulfate is removed and the hyaluronic acid is precipitated, which is anhydrified, dried and ground. Said hyaluronic acid in the form of sodium salt has a minimum richness of 90%, determined by glucuronic acid content, and an average molecular weight between 800,000 and 1,000,000 daltons.

Dermatan sulfate of the compositions of the present invention can be obtained from tissues of birds or mammals, for example from swine or bovine mucosa, bird crests or mammalian skin, following procedures described in the literature (N Volpi, Anal. Biochem. 218, 382-391 (1994); EP 238994).

Preferably, the dermatan sulfate of the compositions of the present invention is obtained from rooster crests, in the process of obtaining hyaluronic acid. Once the dermatan sulfate is separated as a complex, the complex is broken by ionic strength, precipitated, anhydrified, dried and ground. It is also possible to use the porcine mucosa dermatan sulfate marketed by the company Sigma-Aldrich Química (Reference: C3788). Dermatane sulfate in the form of sodium salt that comes from rooster crests has a minimum richness of 90%, determined by photometric titration, and an average molecular weight between 10,000 and 50,000 daltons.

The collagen hydrolyzate of the compositions of the present invention can be obtained from mammalian skin.

or cockscomb, following procedures described in the literature ("Final Report on the Safety Assessment of Hydrolyzed Collagen", Journal of the American College of Toxicology 4, No. 5, 199-221, Mary Ann Liebert, Inc., Publishers, ( 1985)).

The compositions of the present invention can be prepared by mixing their components in the desired proportions. Thus, compositions containing hyaluronic acid and dermatan sulfate, or hyaluronic acid, dermatan sulfate and collagen hydrolyzate (amino acids and low molecular weight peptides) can be prepared.

The compositions of the present invention can also be obtained directly from bird tissues

or of mammals by an extractive procedure. Thus, for example, one can start from frozen rooster crests, which once chopped are digested with a proteolytic enzyme. Subsequently, the enzyme is deactivated by heating, filtered and precipitated with solvents. It is then filtered, washed and dried. Finally you can proceed to milling. The product obtained comprises hyaluronic acid, dermatan sulfate and collagen hydrolyzate (in the form of amino acids and low molecular weight peptides). By slightly modifying this procedure, compositions comprising hyaluronic acid and dermatan sulfate, but not collagen hydrolyzate, can be obtained. The presence of the three components (HA, DS and CH) or of only two (HA and DS) and the proportion between them will depend on the procedure for obtaining (type of enzyme, temperature, reaction time and purification process).

The composition of the invention comprising hyaluronic acid, dermatan sulfate and collagen hydrolyzate is the one comprising 65% hyaluronic acid (HA), 13.40% dermatan sulfate (DS) and 6.56% hydrolyzate of collagen (CH) (weight ratio HA: DS: CH 1: 0.20: 0.10).

In the present invention, chondroitin glycosaminoglycan sulfate has been used solely for comparative purposes. Chondroitin sulfate has a polymeric structure characterized by a repeating disaccharide, consisting of N-acetyl-D-galactosamine and D-glucuronic acid. The majority of N-acetyl-Dgalactosamine residues are sulfated. It can be obtained by proteolytic enzymatic digestion of cartilaginous tissues of animals, for example tracheas of cattle or pigs or cartilaginous skeletons of fish. Chondroitin sulfate that comes from cartilaginous tissue is found mainly in two isomeric forms, chondroitin 4-sulfate and chondroitin 6-sulfate. The average molecular weight of chondroitin sulfate may vary depending on its origin and / or the method used to obtain it. Generally, it has an average molecular weight between 10,000 and 70,000 daltons.

Chondroitin sulfate in the form of sodium salt used for comparative purposes in Example 1 of the present invention is a commercial product available at www.bioiberica.com with a minimum richness of 98%, determined by photometric titration, and a molecular weight half between 14,000 and 18,000 daltons. It is obtained from cartilage of bovine origin.

To use the compositions of the present invention in the treatment or prevention of overweight, obesity, insulin resistance, type 2 diabetes, fatty liver or dyslipidemia in a mammal, they are formulated in suitable pharmaceutical compositions, using conventional techniques and excipients or vehicles. , as described in Remington: The Science and Practice of Pharmacy 2000, edited by Lippincott Williams and Wilkins, 20th edition, Philadelphia. The pharmaceutical compositions comprise a therapeutically effective amount of a composition of the present invention and at least one pharmaceutically acceptable excipient for administration to the patient. Said pharmaceutical compositions can be administered to the patient in required doses. The administration of the pharmaceutical compositions can be carried out by different routes, for example, oral, intravenous, subcutaneous, intramuscular, sublingual, intradermal, nasal or topical. The pharmaceutical compositions of the invention include a therapeutically effective amount of a composition of the present invention, said amount depending on many factors, such as, for example, the physical condition of the patient, age, sex, route of administration, frequency of administration or severity of the illness. Furthermore, it will be understood that said dosage of the composition of the invention can be administered in single or multiple dose units to provide the desired therapeutic effects.

The pharmaceutical compositions of the invention will generally be in solid, liquid or gel form. Pharmaceutical preparations in solid form that can be prepared in accordance with the present invention include powders, mini-granules (pellets), tablets, dispersible granules, capsules, seals, tablets, lyophilisates and suppositories. Liquid form preparations include solutions, suspensions, emulsions, syrups, elixirs, drinkable vials and herbal teas. Also contemplated are preparations of solid forms that are to be converted, immediately before being used, into preparations in liquid form. Such liquid forms include solutions, suspensions and emulsions.

Brief Description of the Figures

Figure 1 shows the images, under the phase contrast microscope, of mouse embryonic fibroblasts (MEFs) grown for 8 days in growth medium containing: vehicle (water, Control); hyaluronic acid (HA) at final concentration (c.f.) 80 µg / mL; dermatan sulfate (DS) at c.f. 20 µg / mL; or composition of the invention HA80 + DS20 a c.f. 100 µg / mL. Intracellular fat drops refractory to the phase contrast microscope are seen in all cultures, except the one treated with HA80 + DS20. (Magnification x200).

Figure 2 shows the expression levels of messenger RNAs (mRNA) for the PPARgamma (A) and C / EBPalfa (B) adipogenic transcription factors and for the lipogenic fatty acid synthase (FAS) protein (C) in MEFs grown during 8 days in growth medium supplemented with: vehicle (water); HA to c.f. 80 µg / mL; DS a c.f. 20 µg / mL; CS to c.f. 20 µg / mL or composition of the invention HA80 + DS20 at c.f. 100 µg / mL. The mRNA levels for beta-actin were used as internal control. The results are expressed in both the level of expression in the vehicle-treated control cells, and are the mean ± SEM of at least six samples for each condition tested, distributed in two independent experiments. ***; P = 0.001 vs vehicle, ttest.

Figure 3 shows the expression levels of messenger RNAs (mRNA) for PPARgamma (A) and C / EBPalfa (B) adipogenic transcription factors and for FAS (C) lipogenic protein in MEFs after 8 days of differentiation between containing an adipogenic hormonal cocktail supplemented with: vehicle (water); HA to

c.f. 80 µg / mL; DS a c.f. 20 µg / mL; or composition of the invention HA80 + DS20 a c.f. 100 µg / mL. The mRNA levels for beta-actin were used as internal control. The results are expressed both by one of the level of expression in the control cells treated with vehicle, and are the mean ± SEM of triplicates for each treatment. *; P <0.05 vs vehicle, t-test.

Figure 4 shows the levels of resistin expression in MEFs after 8 days of differentiation in adipogenic medium in the presence of different final concentrations of HA + DS in a HA to DS weight ratio of 1: 0.25. The mRNA levels for beta-actin were used as internal control. The results are expressed both by one of the level of expression in the control cells treated with vehicle, and are the mean ± SEM of four experiments performed in triplicates. The different letters (a, b) imply significant changes according to ANOVA of a factor (P <0.05).

Figure 5 shows the cumulative intake after 143 days of oral supplementation with vehicle (water, Control group), or with a composition of the invention, at doses 0.45 mg / mouse / day (group HA + DS + CH) . The data represented correspond to the mean ± SEM of 8 animals per group, distributed in 2 cages of 4 animals each. **; P <0.01 HA + DS + CH vs Control, t-test.

Figure 6 shows the circulating glucose levels (A), circulating insulin levels (B) and HOMA-IR index

(C) in mice treated orally with vehicle (water, Control group) or with a composition of the invention, at doses 0.45 mg / mouse / day (HA + DS + CH group). The analyzes were carried out after 88 days of treatment, after the animals were subjected to a 6 h fast. The data correspond to the mean ± SEM of 7-8 animals per group. *; P <0.05 HA + DS + CH vs Control, t-test.

Figure 7 shows the circulating levels of triacylglycerols in the fed state in mice after 143 days of treatment with vehicle (Control group) or with a composition of the invention, at doses 0.45 mg / mouse / day (group HA + DS + CH). The data correspond to the mean ± SEM of 7-8 animals per group. *; P <0.05 HA + DS + CH vs Control, t-test.

Figure 8 shows the evolution of the loss of fat mass (A) and the relative gain of lean mass (B) in mice with dietary obesity in thinning phase, treated daily orally with vehicle (Control group) or with a composition of the invention, at doses 3 mg / mouse / day (group HA + DS + CH). The results are the mean ± SEM of 7-8 animals per group. *; P <0.05 HA + DS + CH vs Control in respective time, t-test.

Figure 9 shows the weight of deposits of inguinal white adipose tissue (TABi), retroperitoneal (TABr) and epididymal (TABe), and body adiposity index in mice exposed for 32 days to thinning conditions and treated with vehicle (Control group ) or with a composition of the invention, at doses 3 mg / mouse / day (group HA + DS + CH). The adiposity index was defined, for each animal, as the sum of the weight of its TAB deposits expressed as a percentage of body weight. The results are the mean ± SEM of 7 animals per group. *; P <0.05 HA + DS + CH vs Control, t-test.

Figure 10 shows the circulating levels in the fed state of glucose (A) and insulin (B) in mice exposed for 32 days to slimming conditions and treated with vehicle (Control group) or with a composition of the invention, at doses 3 mg / mouse / day (group HA + DS + CH). The results are the mean ± SEM of 6-7 animals per group. *; P <0.05 HA + DS + CH vs Control, t-test.

Detailed description of the preferred embodiments:

The following examples are by way of illustration, and do not represent a limitation of the scope of the present invention.

Example 1: Study of the efficacy of a composition of the present invention in the inhibition of spontaneous adipogenesis of mouse embryonic fibroblasts, compared to hyaluronic acid, dermatan sulfate and chondroitin sulfate

Adipogenesis is the process of formation of fat storage cells (adipocytes) from precursor cells. Obesity is often correlated with the presence of an excess of adipocytes in fatty deposits. The objective of the study was to compare the potential of a combination of hyaluronic acid and dermatan sulfate, in a HA to DS weight ratio of 1: 0.25 (HA80 + DS20), and of its individual components of inhibiting intracellular accumulation of Lipids and adipocyte marker gene expression in cultures of primary mouse embryonic fibroblasts (MEFs) maintained in growth medium. A comparison was also made with the chondroitin glycosaminoglycan sulfate. MEFs are multipotent, cultivable cells, capable of differentiating into different mesenchymal cell types (adipocytes, chondrocytes, osteoblasts) depending on the hormonal stimulation they receive. Some studies have reported that serum from the culture medium can induce spontaneous adipogenesis in multipotent mesenchymal cells such as MEFs (L. Wu et al., J. Cell Mol. Med. 14, 922-932 (2010)).

Materials and methods

MEFs obtained from embryos of 13 days of gestational age were grown in monolayer until they reached the confluence, in the middle of growth consisting of AmnioMAXTM-C100 basal medium (Invitrogen, Carlsbad, CA, USA) supplemented with AmnioMAXTM-C100 supplement (7 , 5%), fetal bovine serum (7.5%) and antibiotics, in 12-well culture plates of 1 mL capacity / well, following a previously described protocol (JB Hansen et al., Proc. Natl. Acad. Sci. USA 101, 4112-4117 (2004)). Two days after the confluence was reached, which is considered on day 0 of culture, the cells were exposed to: vehicle (water); hyaluronic acid (HA) at final concentration (c.f.) 80 µg / mL; dermatan sulfate (DS) at c.f. 20 µg / mL; Chondroitin Sulfate (CS) at c.f. 20 µg / mL; or composition of the present invention HA80 + DS20 a c.f. 100 µg / mL, which contained per mL 80 µg of HA and 20 µg of DS. The culture medium in contact with the cells was changed every two days (days 2, 4 and 6 of culture) by fresh medium, always with the supplements indicated above. The cells were collected on day 8 of culture, after being examined and photographed under a phase contrast microscope to visualize their shape and eventual accumulation of lipids. The mRNA levels of the genes related to adipocyte differentiation PPARgamma, C / EBPalfa and fatty acid synthase (FAS) were determined by real-time PCR, from total RNA extracted from the cells with Trizol reagent and retrotranscribed with reverse transcriptase, and using the beta-actin mRNA as an internal control. The pairs of specific primers of each gene were determined using Primer 3 software (Whitehead Institute for Biomedical Research, Cambridge, MA, USA), and their specificity was contrasted by means of the ENTREZ and BLAST databases (National Center for Biotechnology Information, Bethesda , MD, USA). The primers were produced by Sigma (Madrid, Spain). The statistical significance of differences with respect to vehicle treatment was analyzed by Student's t. The threshold of significance was set at P <0.05.

Results

The observation of the cells at day 8 of culture revealed the presence of numerous islands of spontaneously differentiated cells in fat-filled adipocytes, refractory to the phase contrast microscope, in vehicle-treated cultures, HA80 and DS20, but not in those treated with the composition of the invention HA80 + DS20 (Figure 1).

According to the above, at the molecular level, the expression of the adipocyte marker genes PPARy, C / EBPa and FAS was drastically and significantly reduced in the cells treated with the composition of the invention HA80 + DS20, but not in those treated with the components of the composition separately or with CS (Figure 2).

These results indicate that the combination of hyaluronic acid and dermatan sulfate inhibits spontaneous adipogenesis in MEFs in culture in a synergistic effect, which exceeds the sum of the effects of the individual components of the combination separately. Therefore, the composition of the invention can be very useful in the treatment or prevention of overweight and obesity, as well as in reducing fat formation.

Example 2: Study of the efficacy of a composition of the present invention in inhibiting the expression of adipocyte markers during adipogenesis of hormonally induced mouse embryonic fibroblasts, compared to hyaluronic acid and dermatan sulfate

The objective of the study was to compare the potential of a combination of hyaluronic acid and dermatan sulfate, in a HA to DS weight ratio of 1: 0.25 (HA80 + DS20), and of its individual components of affecting adipogenesis in cultures of mouse embryonic fibroblasts (MEFs) hormonally stimulated to differentiate into adipocytes.

Materials and methods

MEFs obtained from embryos of 13 days of gestational age were grown in monolayer until they reached the confluence in 12-well culture plates of 1 mL capacity / well, as described for Example 1. Two days later Once the confluence was reached (day 0), the cells were stimulated by exposing them for 48 hours to a standard adipogenic hormonal cocktail containing dexamethasone, methyl isobutylxanthine, insulin and rosiglitazone. Subsequently, the cells received fresh growth medium supplemented with insulin and rosiglitazone every two days (days 2, 4 and 6). The differentiation process was carried out in cells exposed from day 0 to: vehicle (water); HA at final concentration (c.f.) 80 µg / mL; DS a c.f. 20 µg / mL; or composition of the present invention HA80 + DS20 a c.f. 100 µg / mL, which contained per mL 80 µg of HA and 20 µg of DS. The cells were collected on day 8 of culture. The mRNA levels of selected genes were determined by real-time PCR, as described for Example 1.

Results

The expression of the adipocyte marker genes PPARgamma, C / EBPalfa and FAS was reduced in the cells exposed to the composition of the invention HA80 + DS20 during the adipogenic process, but not in those exposed to the individual components of the composition, HA or DS, separately (Figure 3).

These results indicate that the combination of HA and DS exerts a synergistic anti-adipogenic effect even under environmental conditions (presence of a pro-adipogenic hormonal cocktail) that promote this process. Consequently, the composition of the invention may be useful in the treatment or prevention of overweight and obesity, as well as in reducing fat formation.

Example 3: Study of the efficacy of a composition of the present invention in inhibiting the expression of resistin, an adipokine related to insulin resistance, in adipocytes derived from MEFs

The objective was to study the effect of a combination of hyaluronic acid and dermatan sulfate, at different concentrations, in a HA to DS weight ratio of 1: 0.25 (HA + DS), on the expression of secretion proteins of the adipocytes that have been previously implicated as factors that enhance insulin resistance, particularly resistin, in fat cells in culture. In the same study, the effect of hyaluronic acid and dermatan sulfate was also determined separately.

Materials and methods

MEFs obtained from embryos of 13 days of gestational age were grown in monolayer until they reached confluence and exposed to a standard adipogenic hormonal cocktail as described for Example 2, in the presence of: vehicle (water); HA to c.f. 80 µg / mL; DS a c.f. 20 µg / mL; Composition of the invention HA16 + DS4 a c.f. 20 µg / mL, which contained per mL 16 µg of HA and 4 µg of DS; composition of the invention HA80 + DS20 a c.f. 100 µg / mL, which contained per mL 80 µg of HA and 20 µg of DS; or composition of the invention HA160 + DS40 a c.f. 200 µg / mL, which contained per mL 160 µg of HA and 40 µg of DS. The cells were collected on day 8 of culture. Resistance gene expression levels were determined by real-time PCR, as described for Example 1.

Results

Exposure to the compositions of the invention HA16 + DS4 at final concentration 20 µg / mL, HA80 + DS20 at final concentration 100 µg / mL or HA160 + DS40 at final concentration 200 µg / mL, reduced resistin expression after 8 days of adipocyte differentiation of MEFs, in a dose-dependent manner (Figure 4). The effect of the compositions of the invention on the gene expression of resistin was not reproduced by the individual components HA80 and DS20 when they were tested separately. Accordingly, the compositions of the invention may be useful in the treatment or prevention of insulin resistance and type 2 diabetes, as well as in increasing insulin sensitivity.

Example 4: Effect of supplementation with a composition of the present invention on the reduction of weight gain and energy intake in mice

The objective was to study the effect of oral supplementation with a composition of the present invention (HA + DS + CH) on the evolution of biometric parameters in vivo, in mice.

A composition containing 65% hyaluronic acid (HA), 13.40% dermatan sulfate (DS) and 6.56% collagen hydrolyzate (CH) was used (weight ratio HA: DS: CH 1: 0 , 20: 0.10).

Materials and methods

Male C57BL6 / J mice (Charles River Laboratories, Barcelona, Spain) of 6 weeks of age at the beginning of the experiment were set at 22 ° C with a light / dark period of 12 hours, under ad libitum feeding conditions with a defined standard diet ( manufactured by Research Diets, Inc, New Brunswick, NJ, USA) containing 3.85 Kcal / g of diet. At the start of the experiment, two experimental groups were established (n = 8 animals per group), which they received orally, for 143 days, vehicle (water, Control group), or a composition of the present invention (HA + DS + CH ), at a dose of 0.45 mg / mouse / day (group HA + DS + CH; weight ratio HA: DS: CH 1: 0.20: 0.10). The body weight and intake of the animals were monitored during the 143 days of treatment. The statistical significance of the observed effects was analyzed by Student's t test. The threshold of significance was set at P <0.05.

Results

Animals treated with the composition of the invention HA + DS + CH tended to gain less weight and, at the end of the experiment, their body weight was 5% lower than that of the controls (mean ± SEM: initial weights: Control group, 22.5 ± 0.8 g; HA + DS + CH group, 22.5 ± 0.7 g; final weights: Control group, 29.0 ± 1.0 g; HA + DS + CH group, 27.5 ± 1.0 g).

Cumulative intake during the 143 days of the experiment was 11% lower in animals treated with HA + DS + CH than in animals in the Control group to which the vehicle was administered (P <0.05) (Figure 5).

The results obtained in vivo allow us to affirm that the compositions of the present invention can be useful in the treatment or prevention of overweight and obesity, in reducing body weight, weight gain, food intake and appetite, in preventing weight gain and induce the feeling of satiety.

Example 5: Effect of oral supplementation with a composition of the present invention on parameters related to insulin sensitivity in mice

The objective was to study the effect of oral supplementation with a composition of the present invention (HA + DS + CH) on insulin sensitivity in vivo, in mice.

A composition containing 65% hyaluronic acid (HA), 13.40% dermatan sulfate (DS) and 6.56% collagen hydrolyzate (CH) was used (weight ratio HA: DS: CH 1: 0 , 20: 0.10).

Materials and methods

The same experimental groups as in Example 4 were used. After 88 days of treatment, the animals of both groups, Control and HA + DS + CH, were subjected to a 6 h fast after which blood was taken, for the determination of circulating glucose levels (by Roche's Accu-Check system) and insulin (using an ELISA kit, distributed by DGR Instruments GnbH). For each animal, the HOMA-IR index (homeostatic model assessment for insulin resistance) was calculated, which is a well-accepted index indicative of insulin relative sensitivity, using the formula of Matthews et al. (D.R. Matthews et al., Diabetologia 28, 412-419 (1985)): HOMA-IR = fasting glucose (mM) x fasting insulin (mU / L) / 22.5. The lower the HOMA-IR index, the lower the insulin resistance and therefore the greater the sensitivity to this hormone. This index has been validated in humans and mice (S. Lee et al. Am. J. Physiol. Endocrinol. Metab. 294, E261-70 (2008)).

Results

Animals supplemented with HA + DS + CH at the same blood glucose showed lower insulinemia than control animals, and therefore a HOMA-IR index also lower (Figure 6). These results indicate that insulin sensitivity is higher in the group supplemented with the composition of the invention comprising hyaluronic acid, dermatan sulfate and collagen hydrolyzate (HA + DS + CH) than in the control group. Impaired insulin sensitivity is considered one of the main triggers for the development of type 2 diabetes, along with pancreatic beta cell dysfunction (D. LeRoith, Am. J. Med. 113 (Suppl 6A), 3S -11S (2002)).

The in vivo results obtained in the present study make it possible to affirm that the compositions of the present invention may be useful in the treatment or prevention of type 2 diabetes and insulin resistance and in increasing insulin sensitivity.

Example 6: Effect of oral supplementation with a composition of the present invention on the levels of triacylglycerols in mice

The objective was to study the effect of oral supplementation with a composition of the present invention (HA + DS + CH) on circulating levels of triacylglycerols in vivo, in mice.

A composition containing 65% hyaluronic acid (HA), 13.40% dermatan sulfate (DS) and 6.56% collagen hydrolyzate (CH) was used (weight ratio HA: DS: CH 1: 0 , 20: 0.10).

Materials and methods

The same experimental groups as in Example 4 were used. After 143 days of treatment, the animals of both groups, Control and HA + DS + CH, were sacrificed by decapitation. Circulating serum triglyceride levels prepared from neck blood were determined by a commercial colorimetric enzymatic assay (Sigma).

Results

After 143 days of supplementation, the animals supplemented with the composition of the invention comprising hyaluronic acid, dermatan sulfate and collagen hydrolyzate (HA + DS + CH) had lower triglyceridemia than the controls to which the vehicle had been administered (Figure 7). Consequently, the compositions of the present invention can be of great utility in the treatment or prevention of dyslipidemia and in reducing blood triglycerides.

Example 7: Effect of oral supplementation with a composition of the present invention on the reversal of obesity in mice

The objective was to study weight and fat loss in obese mice when treated orally with a composition of the present invention (HA + DS + CH).

A composition containing 65% hyaluronic acid (HA), 13.40% dermatan sulfate (DS) and 6.56% collagen hydrolyzate (CH) was used (weight ratio HA: DS: CH 1: 0 , 20: 0.10).

Materials and methods

25-week-old male C57BL6 / J mice were employed who had previously been induced to dietary obesity by feeding with a hyperlipid diet from 6 weeks of age. The mice had, at the time of the start of the experiment, 22% excess weight compared to animals of the same age maintained under a standard diet. At the beginning of the experiment, the obese mice became fed with a normolipid diet, and were distributed in two groups, which received daily, orally, vehicle (water, 60 µl, Control group) or a composition of the present invention (HA + DS + CH), at a dose of 3 mg / mouse / day (n = 7-8 animals / group) (group HA + DS + CH; weight ratio HA: DS: CH 1: 0.20: 0.10). In both groups, Control and HA + DS + CH, body weight was monitored regularly, as well as body composition using a non-invasive magnetic resonance system (ECHO-MRI). The animals were sacrificed by decapitation, 32 days after the start of the standard diet reversal, proceeding with the collection of blood and tissues, and the weight of adipose tissues. The statistical significance of the observed effects was analyzed by the Student t test. The threshold of significance was set at P <0.05.

Results

Animals treated with HA + DS + CH tended to lose more weight percentage and faster than those in the Control group; this trend reached statistical significance after 11 days of reversion to the normolipid diet, at which point the animals in the Control group had lost 9.5% of their initial body weight, and the animals treated with HA + DS + CH, 12, 8% In addition, the animals treated with HA + DS + CH had a greater and faster percentage loss of fat mass than the animals of the Control group (Figure 8A), which correlated with a greater relative increase in their lean mass (Figure 8B) .

At the end of the experiment, body adiposity (sum of the weights of adipose tissue deposits as a percentage of body weight) of animals treated with HA + DS + CH was 30% lower than that of animals in the Control group ( Figure 9B). All dissected fatty deposits (inguinal (TABi), retroperitoneal (TABr) and epididymal (TABe)) were significantly smaller in animals treated with HA + DS + CH (Figure 9A).

At the end of the experiment, animals treated with HA + DS + CH, despite not showing variations in blood glucose, showed lower insulinemia than Control animals, indicative of greater insulin sensitivity (Figure 10).

These results obtained in vivo allow us to affirm that the compositions of the present invention can be useful in the treatment or prevention of overweight, obesity, type 2 diabetes, fatty liver and insulin resistance, and in reducing body fat, fat formation and body weight.

Claims (22)

1.- Use of a composition comprising hyaluronic acid and dermatan sulfate for the preparation of a medicament for the treatment or prevention of at least one of the diseases that make up the metabolic syndrome in a mammal, selected from the group formed by overweight, obesity , insulin resistance, type 2 diabetes, fatty liver and dyslipidemia. 2. Use according to claim 1 for the preparation of a medicament for the treatment or prevention of overweight associated with metabolic syndrome in a mammal. 3. Use according to claim 1 for the preparation of a medicament for the treatment or prevention of obesity associated with the metabolic syndrome in a mammal. 4. Use according to claim 1 for the preparation of a medicament for the treatment or prevention of insulin resistance associated with metabolic syndrome in a mammal. 5. Use according to claim 4, wherein insulin resistance is associated with overweight or obesity. 6. Use according to claim 1 for the preparation of a medicament for the treatment or prevention of type 2 diabetes associated with the metabolic syndrome in a mammal. 7. Use according to claim 5, wherein type 2 diabetes is associated with overweight or obesity. 8. Use according to claim 1 for the preparation of a medicament for the treatment or prevention of fatty liver associated with the metabolic syndrome in a mammal. 9. Use according to claim 8, wherein the fatty liver is associated with overweight or obesity. 10. Use according to claim 1 for the preparation of a medicament for the treatment or prevention of dyslipidemia associated with metabolic syndrome in a mammal. 11. Use according to claim 10, wherein the dyslipidemia is associated with overweight or obesity. 12. Use according to claim 10 or 11, wherein the dyslipidemia is hypercholesterolemia and hypertriglyceridemia. 13. Use according to claim 10 or 11, wherein the dyslipidemia is hypercholesterolemia. 14. Use according to claim 10 or 11, wherein the dyslipidemia is hypertriglyceridemia. 15. Use according to any one of claims 1 to 14, wherein the composition comprises hyaluronic acid and dermatan sulfate present in a weight ratio hyaluronic acid to dermatan sulfate between 1: 0.05 and 1: 0, 75, preferably between 1: 0.10 and 1: 0.50. 16. Use according to claim 15, wherein the composition comprises hyaluronic acid and dermatan sulfate present in a weight ratio hyaluronic acid to dermatan sulfate of 1: 0.25. 17. Use according to any one of claims 1 to 16, wherein the composition further comprises collagen hydrolyzate. 18. Use according to claim 17, wherein the composition comprises hyaluronic acid, dermatan sulfate and collagen hydrolyzate present in a weight ratio of hyaluronic acid to dermatan sulfate to collagen hydrolyzate between 1: 0.06: 0 , 03 and 1: 0.80: 0.40, preferably between 1: 0.10: 0.05 and 1: 0.40: 0.20. 19. Use according to claim 18, wherein the composition comprises hyaluronic acid, dermatan sulfate and collagen hydrolyzate present in a weight ratio of hyaluronic acid to dermatan sulfate to collagen hydrolyzate of 1: 0.20: 0, 10. 20. Use according to any one of claims 1 to 19, wherein the mammal is a human. 21. Use according to any one of claims 1 to 20, wherein the medicament is adapted for oral administration. X200 FIGURE 1

TO)

1.25

1.00

PPAR mRNA / / -actin mRNA

0.50 0.75

0.25

0.00

vehicle HE HAS DS CS HA80 + DS20

B)

1.25

1.00

C / EBPa mRNA / / -actin mRNA

0.50 0.75

0.25

0.00

vehicle HE HAS DS CS HA80 + DS20

C)

1.50

1.25

FAS mRNA / / -actin mRNA

0.50 0.75 1.00

0.25

0.00

vehicle HE HAS DS CS HA80 + DS20

FIGURE 2

TO) 1.40 1.20 1.00  PPARy mRNA / 0.80 / -actin mRNA   0.60 0.40 0.20 0.00 B) 1.20 1.00 0.80  C / EBPa mRNA / 0.60 / -actin mRNA 0.40 0.20  0.00 C) 1.75 1.50 1.25 1.00  FAS mRNA / / -actin mRNA 0.75 0.50 0.25 0.00  vehicle HA DS HA80 + DS20 FIGURE 3 mRNA resistin / -actin mRNA

1 1.2

0 0.2 0.4 0.6 0.8 0

25 fifty 75 100 125 150 175 200 225

Final concentration (! G / mL) of HA + DS (weight ratio 1: 0.25)

 cumulative intake (Kcal / animal) FIGURE 4 1700 1600 1500 1400 1300 1200 1100 1000 HA + DS + CH control FIGURE 5  A) 8  7 6 circulating glucose5 4 (mM) 3 2 1 0 B) 1.2 1.0 0.8  circulating insulin 0.6 (! g / L) 0.4 0.2 0.0 C) 8.0 7.0 6.0 5.0 HOMA-IR 4.0 3.0 2.0 1.0 0.0 HA + DS + CH control HA + DS + CH control  HA + DS + CH control FIGURE 6  triacylglycerol circulators (mg / mL) 15.0 12.5 10.0 7.5 5.0 2.5 0.0 HA + DS + CH control FIGURE 7 A) B) time (days) Loss of fat mass (%) P = 0.132 12 10 4 2 0 -10 time (days) FIGURE 8 TO)  TABi TABr TABe B) 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0  HA + DS + CH control FIGURE 9 Adiposity (%) TO) B) circulating insulin (! g / L) circulating glucose (mM)  10 9 8 7 6 5 4 3 2 1 0 Control 2.5 2.0 1.5 1.0 0.5  0.0 Control FIGURE 10 HA + DS + CH  HA + DS + CH SUMMARY The present invention relates to a composition comprising hyaluronic acid and dermatan sulfate for use in the treatment or prevention of at least one of the diseases that make up the metabolic syndrome in a mammal, selected from the group consisting of overweight, obesity, resistance to insulin, type 2 diabetes, fatty liver or dyslipidemia. In addition the composition may contain collagen hydrolyzate. The composition is in the form of a pharmaceutical composition. TO)  TABi TABr TABe B) Adiposity (%) 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0  HA + DS + CH control FIGURE 9 SPANISH OFFICE OF THE PATENTS AND BRAND Application no .: 201231198 SPAIN Date of submission of the application: 25.07.2012 Priority Date: REPORT ON THE STATE OF THE TECHNIQUE 51 Int. Cl.: See Additional Sheet RELEVANT DOCUMENTS

Category

56 Documents cited Claims Affected

TO

JP 2008 150326 A (GLOBE COMPANY INC) 03.07.2008, (summary) Derwent Publications Ltd., AN 2008-J12845. 1-21

TO

WO 2011130261 A1 (RUFF KEVIN) 20.10.2011, the whole document. 1-21

TO

WO 2007059874 A2 (BIOIBERICA S.A.) 05/31/2007, the whole document. 1-21

TO

US 2005084518 A1 (ARAI) 21.04.2005, the whole document. 1-21

Category of the documents cited X: of particular relevance Y: of particular relevance combined with other / s of the same category A: reflects the state of the art O: refers to unwritten disclosure P: published between the priority date and the date of priority submission of the application E: previous document, but published after the date of submission of the application

This report has been prepared • for all claims • for claims no:

Date of realization of the report 18.12.2013

Examiner M. Cumbreño Galindo Page 1/4

REPORT OF THE STATE OF THE TECHNIQUE Application number: 201231198 CLASSIFICATION OBJECT OF THE APPLICATION A61K31 / 728 (2006.01) A61K31 / 737 (2006.01) A61P3 / 00 (2006.01) Minimum documentation sought (classification system followed by classification symbols) A61K, A61P Electronic databases consulted during the search (name of the database and, if possible, terms of search used) INVENES, EPODOC, WPI, MEDLINE, NPL, EMBASE, BIOSIS XPESP, XPESP2, XPOAC State of the Art Report Page 2/4 WRITTEN OPINION Application number: 201231198 Date of Completion of Written Opinion: 18.12.2013 Statement

Novelty (Art. 6.1 LP 11/1986)

Claims Claims 1-21 IF NOT

Inventive activity (Art. 8.1 LP11 / 1986)

Claims Claims 1-21 IF NOT

The application is considered to comply with the industrial application requirement. This requirement was evaluated during the formal and technical examination phase of the application (Article 31.2 Law 11/1986). Opinion Base.- This opinion has been made on the basis of the patent application as published. State of the Art Report Page 3/4 WRITTEN OPINION Application number: 201231198 1. Documents considered.- The documents belonging to the state of the art taken into consideration for the realization of this opinion are listed below.

Document

Publication or Identification Number publication date

D01

JP 2008 150326 A 03.07.2008

D02

WO 2011130261 A1 20.10.2011

D03

WO 2007059874 A2 05/31/2007

D04

US 2005084518 A1 04/21/2005

2. Statement motivated according to articles 29.6 and 29.7 of the Regulations for the execution of Law 11/1986, of March 20, on Patents on novelty and inventive activity; quotes and explanations in support of this statement The present invention aims at the use of a composition comprising hyaluronic acid and dermatan sulfate for the preparation of a medicament for the treatment or prevention of at least one of the diseases that make up the metabolic syndrome in a mammal (claims 1 to 21 ). D01 anticipates a drink containing hyaluronic acid, dermatan sulfate, vitamins, marine collagen and other compounds, as well as its use in the prevention of diseases such as diabetes, hypercholesterolemia or triglyceridemia. D02 discloses a method to treat a disease related to abnormal glucose metabolism, such as metabolic syndrome, which consists of orally administering eggshell-derived material. Said material consists of hexosamine, glycosaminoglycans such as dermatan sulfate, hyaluronic acid or collagen, among other ingredients. D03 discloses the use of compositions comprising hyaluronic acid and dermatan sulfate for the preparation of oral medications for the treatment of osteoarthritis. The compositions may also contain collagen hydrolyzate and nucleic acids and may also be used in food processing or nutritional supplements. D04 anticipates foods containing hyaluronic acid and dermatan sulfate and its use to improve the appearance of the skin and reduce hair loss. NEW AND INVENTIVE ACTIVITY The cited documents disclose other uses of compositions comprising hyaluronic acid and dermatan sulfate, or the use of said compositions for the treatment of diseases associated with abnormal glucose metabolism, such as metabolic syndrome, which includes diabetes or dyslipidemia. In these compositions, hyaluronic acid and dermatan sulfate are mixed with other ingredients without making clear the role of both compounds in relation to the aforementioned treatment, or they are possible constituents of the composition, although there seems to be indications of their use in formulations intended for the treatment of diseases that make up the metabolic syndrome. Therefore, claims 1 to 21 can be considered new in view of the state of the art and have inventive activity. State of the Art Report Page 4/4